Signosis

Product Name	Catalog #	Price		Qty
NFkB Luciferase Reporter Jurkat Stable Cell Line	SL-0050-FP	$3,870	X	- +

Description(s):

NFkB Luciferase Reporter Jurkat Stable Cell Line is derived from human T lymphocyte cells, and stably express firefly luciferase reporter gene under the control of the NF-kB response enhancer element. This cell line is an ideal cellular model for monitoring the activation of NFkB pathway triggered by stimuli treatment, enforced gene expression and gene knockdown. This cell line express CD3 and CD28 (see data below).

Principle

NF-κB is a critical regulator of inflammatory responses, proliferation and differentiation of T-cells. The aberrant activation of NFkB can contribute to the development of autoimmunity, chronic inflammation, or lymphoid cancer. Jurkat cells are human T lymphocyte cells widely used to study T cell signaling. Signosis developed a stable Jurkat NFkB-luciferase reporter stable cell line, which can be used for easily monitoring the activation of NFkB activation in T cells through sensitive luciferase analysis. This cell line was established by transfection using a pTA-NFkB-luciferase reporter vector, along with hygromycin expression vector followed by hygromycin selection. The hygromycin resistant clones were subsequently screened for luciferase activity induced by TNFapha treatment.

Principle behind TF luciferase reporter. TF luciferase reporter stable cell line utilizes artificial promoter constructs to drive luciferase expression. The promoter region can consists of multiple repeats of a cis-element TF binding site, a DNA fragment from the promoter region of a known TF downstream gene, or a DNA fragment containing putative/known TF binding sites. There are several ways that a TF can be activated, such as through extracellular stimuli or through intracellular signaling pathways. Once activated, the TF translocates to the nucleus and often interacts with relevant co-factors to drive gene expression. Once luciferase is expressed, it can generate light in an enzymatic assay and the amount of light measured is positively correlated with the level of TF activation.

Data

Analysis of SL-0050 NFKB Luciferase Reporter Jurkat T Stable Cell Line. 0.1x10⁶Cells were plated on a 96-well plate and incubate in 100μl media +0.1% FBS . Additions to wells were as follows: (Control ) No addition; (PHA) 50μg/ml incubated for 20 hours; (PHA+TNFα) PHA50μg/ml incubated for 4 hours then TNFα 20ng/ml was added and continue incubating for 16 hours, (TNFα) TNFα 20ng/ml incubated for 16 hours.

SL-0050 luc activity upon induction with CD3 and CD28 Abs. These cell line express CD3 and CD28 and can be activated by exposure to CD3 or CD28 antibodies or combination of both. 0.1x10⁶Cells were plated on a 96-well plate and incubate in 100μl media +0.1% FBS . CD3 antibody (1ug/ml) and CD28 antibody (2ug/ml) were added to the cells and incubated for 16 hours.

All data was read on a luminometer with a sensitivity of 3x10^-21 moles luciferase.

Literature

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Citations

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Withaferin A induces Nrf2-dependent protection against liver injury: role of Keap1-independent mechanisms. DL Palliyaguru, DV Chartoumpekis, N Wakabayashi. Free Radical Biology and Medicine 2016. http://dx.doi.org/10.1016/j.freeradbiomed.2016.10.003

Occurrence and in vitro toxicity of unregulated disinfection by-products in two Saskatchewan drinking water treatment plants. Christena Leigh Watts. Handle Proxy, Universidade Da Coruña, 1 Sept. 2018, hdl.handle.net/10388/11499.

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Redox-responsive, reversibly-crosslinked thiolated cationic helical polypeptides for efficient siRNA encapsulation and delivery.N Zheng, Z Song, Y Liu, R Zhang, R Zhang, C Yao. Journal of Controlled Release.Volume 205, 10 May 2015, Pages 231–239

Targeted delivery of cisplatin to tumor xenografts via the nanoparticle component of nano-diamino-tetrac. Thangirala Sudha, Dhruba J Bharali, Murat Yalcin, Noureldien HE Darwish, Melis Debreli Coskun, Kelly A Keating, Hung-Yun Lin, Paul J Davis, Shaker A Mousa. February 2017 ,Vol. 12, No. 3, Pages 195-205 , DOI 10.2217/nnm-2016-0315.

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Thymoquinone Modulates Blood Coagulation in Vitro via Its Effects on Inflammatory and Coagulation Pathways. V Muralidharan-Chari, J Kim, A Abuawad, M Naeem. Int. J. Mol. Sci. 2016, 17(4), 474; doi:10.3390/ijms17040474

Drug repurposing screen identifies lestaurtinib amplifies the ability of the poly (ADP-ribose) polymerase 1 inhibitor AG14361 to kill breast cancer associated gene-1 …G Vazquez-Ortiz, C Chisholm, X Xu, TJ Lahusen, C Li… - Breast Cancer Research 2014, Jun 24;16(3):R67. doi: 10.1186/bcr3682.

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Alcohol promotes migration and invasion of triple-negative breast cancer cells through activation of p38 MAPK and JNK†M Zhao, EW Howard, AB Parris, Z Guo, Q Zhao, X Yang. Molecular Carcinogenesis, 2016, DOI: 10.1002/mc.22538

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Active β-catenin is regulated by the PTEN/PI3 kinase pathway: a role for protein phosphatase PP2A. Amit Persad,1,* Geetha Venkateswaran,1,* Li Hao,1 Maria E. Garcia,1 Jenny Yoon,1 Jaskiran Sidhu,1 and Sujata Persad1. Genes Cancer. 2016 Nov; 7(11-12): 368–382., doi: 10.18632/genesandcancer.128

Diallyl trisulfide inhibits proliferation, invasion and angiogenesis of glioma cells by inactivating Wnt/β-catenin signaling. Qingxia Tao, Cuiying Wu, Ruxiang Xu, Lijun Niu, Jiazhen Qin, Ning Liu, Peng Zhang, Chong Wang. Cell and Tissue Research, 2017, doi:10.1007/s00441-017-2678-9.

BCN057 induces intestinal stem cell repair and mitigates radiation-induced intestinal injury. Payel Bhanja, Andrew Norris, Pooja Gupta-Saraf, Andrew Hoover and Subhrajit Saha. Stem Cell Research & Therapy, vol. 9, no. 1, Feb. 2018, doi:10.1186/s13287-017-0763-3.

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Direct Antioxidant Properties of Methotrexate: Inhibition of Malondialdehyde-Acetaldehyde-Protein Adduct Formation and Superoxide Scavenging. Matthew C. Zimmermana, Dahn L. Clemensb, c, Michael J. Duryeeb, Cleofes Sarmientoa, Andrew Chioub, Carlos D. Hunterb, Jun Tiana, Lynell W. Klassenb, c, James R. O’Dellb, c, Geoffrey M. Thieleb, c, Redox Biology,2017.07.018, doi: 10.1016/j.redox.2017.07.018

Active β-catenin is regulated by the PTEN/PI3 kinase pathway: a role for protein phosphatase PP2A. Amit Persad1,*, Geetha Venkateswaran1,*, Li Hao1, Maria E. Garcia1, Jenny Yoon1, Jaskiran Sidhu1 and Sujata Persad1.Genes & Cancer. January 30, 2017 . https://doi.org/10.18632/genesandcancer.128

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Hypothermia augments stress response in mammalian cells. Eskla, Kattri-Liis, et al. Free Radical Biology and Medicine, 2018, doi:10.1016/j.freeradbiomed.2018.04.571.

NFkB Luciferase Reporter HepG2 Stable Cell Line (SL-0017)

Hypothermia augments stress response in mammalian cells. Eskla, Kattri-Liis, et al. Free Radical Biology and Medicine, 2018, doi:10.1016/j.freeradbiomed.2018.04.571.

CHOP Luciferase Reporter Mia_Paca2 Stable Cell Line (SL-0025)

A new ent-norabietant diterpenoid from roots of Euphorbia lathyris L. Yun, Young Sook, et al. Tetrahedron Letters, 2018, doi:10.1016/j.tetlet.2018.06.017.

NFAT Luciferase Reporter Jurkat Stable Cell Line (SL-0032)

Diversification of Bw4 Specificity and Recognition of a Nonclassical MHC Class I Molecule Implicated in Maternal–Fetal Tolerance by Killer Cell Ig-like Receptors of the Rhesus Macaque. Banerjee, Priyankana, et al. The Journal of Immunology, 2018, doi:10.4049/jimmunol.1800494.

P329G-CAR-J: a Novel Jurkat-NFAT-Based CAR-T Reporter System Recognizing the P329G Fc Mutation. Darowski, Diana, et al. Protein Engineering, Design and Selection, 2019, doi:10.1093/protein/gzz027.

Dendritic cells dictate responses to PD-L1 blockade cancer immunotherapy. Mayoux, Maud, et al. Science Translational Medicine Vol. 12, Issue 534, 2020.
DOI: 10.1126/scitranslmed.aav7431

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Innovative drinking water treatment techniques reduce the disinfection-induced oxidative stress and genotoxic activity. Lundqvist, Johan, et al. Water Research, vol. 155, 2019, pp. 182–192., doi:10.1016/j.watres.2019.02.052.

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Discovery of a New STAT3 Inhibitor Acting on the Linker Domain. Koseki, Tatsuya, et al. Biological and Pharmaceutical Bulletin, vol. 42, no. 5, 2019, pp. 792–800., doi:10.1248/bpb.b18-00992.

An engineered ligand trap inhibits leukemia inhibitory factor as pancreatic cancer treatment strategy. Hunter, S.A., McIntosh, B.J., Shi, Y. et al. Commun Biol 4, 452 (2021). https://doi.org/10.1038/s42003-021-01928-2

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Discovery of a New STAT3 Inhibitor Acting on the Linker Domain. Koseki, Tatsuya, et al. Biological and Pharmaceutical Bulletin, vol. 42, no. 5, 2019, pp. 792–800., doi:10.1248/bpb.b18-00992.

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Thiamine and Benfotiamine Protect Neuroblastoma Cells against Paraquat and β-Amyloid Toxicity by a Coenzyme-Independent Mechanism. Sambon, Margaux, et al. Heliyon, vol. 5, no. 5, 2019, doi:10.1016/j.heliyon.2019.e01710.

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Vitale LA, He LZ, Thomas LJ, et al. Development of CDX-527: a bispecific antibody combining PD-1 blockade and CD27 costimulation for cancer immunotherapy. Cancer Immunology, Immunotherapy : CII. 2020 May. DOI: 10.1007/s00262-020-02610-y.

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Targeting the Liver‐Brain Axis with Hop‐Derived Flavonoids Improves Lipid Metabolism and Cognitive Performance in Mice. Paraiso, I.L., Revel, J.S., Choi, J., Miranda, C.L., Lak, P., Kioussi, C., Bobe, G., Gombart, A.F., Raber, J., Maier, C.S. and Stevens, J.F. Mol. Nutr. Food Res. 2020, 2000341. https://doi.org/10.1002/mnfr.202000341

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Glass-bottled drinking water: a time capsule to study the historic presence of hazardous chemicals using effect-based methods. Lundqvist, J., Persson, K.M. & Oskarsson, A. Environ Sci Eur 33, 34 (2021). https://doi.org/10.1186/s12302-021-00476-0